Haohao Huang

Supplementary Table S6 shows the sequences of shRNAs used in this study.

Supplementary Table S3 shows UCSC-predicted transcription factors that bind to CKB promoters.

Supplementary Table S1 shows PCr IHC staining score in human glioma microarray.

Supplementary Table S4 shows PCr and BRD2 IHC staining scores in the serial sections of human glioma microarrays.

Supplementary Figures S1 shows that GSCs produce high levels of phosphocreatine through upregulating CKB. Supplementary Figures S2 shows that ZEB1 promotes CKB transcription in GSCs. Supplementary Figures S3 shows that knockdown of CKB impedes GBM growth. Supplementary Figures S4 shows that disruption of phosphocreatine production impedes GBM growth. Supplementary Figures S5 shows that cCr treatment shows no side effect on mice. Supplementary Figures S6 shows that phosphocreatine binds to BRD2 and inhibits its ubiquitin mediated proteasomal degradation. Supplementary Figures S7 shows that phosphocreatine promotes chromosome segregation and GSC proliferation through BRD2 mediated transcription. Supplementary Figures S8 shows that disruption of phosphocreatine biosynthesis by cCr improves JQ1 therapeutic efficacy in GBM.

Supplementary Table S5 shows the detailed genetic mutations of four GBM cell lines.

Supplementary Table S2 shows CKB IHC staining score in human glioma microarray.

Supplementary Table S7 shows the primer sequences used for Real-Time PCR.

Background: Colorectal cancer (CRC) patients are more susceptible to infections due to immune dysregulation. The interleukin-17A (IL-17A) signaling pathway plays a critical role in both immune defense and cancer metastasis. This study aimed to identify novel microRNAs (miRNAs) involved in IL-17A-mediated CRC progression. Methods: RNA sequencing (RNA-seq) was used to analyze gene expression in stimulated peripheral blood mononuclear cells (PBMCs) and HCT116 cells cultured with their conditioned media (CM). Candidate miRNAs involved in IL-17A or transforming growth factor-β (TGF-β)-driven CRC progression were evaluated by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), western blotting, and wound healing assays. Prognostic relevance was assessed in The Cancer Genome Atlas colon adenocarcinoma (TCGA-COAD) cohort via Cox regression and Kaplan-Meier survival analysis. Results: IL-17 signaling emerged as one of the most significantly enriched pathways following immune stimulation. Both IL-17A and TGF-β upregulated the epithelial-mesenchymal transition (EMT) marker vimentin and promoted CRC cell migration. Among the five candidate miRNAs identified, miR-501-5p was markedly induced by IL-17A and TGF-β. Functional assays revealed that either transient overexpression or inhibition of miR-501-5p significantly altered CRC cell migration in response to IL-17A. Notably, elevated miR-501 expression was associated with more than a twofold increased risk of death in stage IV CRC patients from the TCGA-COAD cohort. Conclusions: IL-17 signaling may drive CRC progression and poor clinical outcomes in part through the induction of miR-501-5p, underscoring its potential role in immune-related cancer metastasis and as a prognostic biomarker.

Background: Glioma stem-like cells (GSCs) are key drivers of treatment resistance and recurrence in glioblastoma (GBM). Phosphoglycerate dehydrogenase (PHGDH), a crucial enzyme in the de novo serine synthesis pathway (SSP), is implicated in tumorigenesis and therapy resistance across various cancers. However, its specific role in GBM, particularly in radioresistance, remains poorly understood. Methods: In silico analysis of GBM patient data assessed SSP enrichment and PHGDH expression linked with tumor stemness. Comparative gene expression analysis focused on PHGDH in paired GBM specimens and GSCs. Genetic and pharmacological loss-of-function assays were performed in vitro and in vivo to evaluate PHGDH’s impact on GSC self-renewal and malignant progression. Comprehensive transcriptomic and metabolomic analyses, along with chromatin immunoprecipitation, mass spectrometry, and various other biochemical assays, were used to elucidate PHGDH-mediated mechanisms in GBM progression and radioresistance. Results: PHGDH expression is significantly elevated in GSCs, associated with aggressive glioma progression and poor clinical outcomes. PHGDH activation enhances GSC self-renewal by regulating redox homeostasis, facilitating one-carbon metabolism, and promoting DNA damage response via SSP activation. Importantly, MYC was identified as a crucial transcriptional regulator of PHGDH expression. Furthermore, genetic ablation or pharmacological inhibition of PHGDH markedly reduced tumor growth and increased tumor sensitivity to radiotherapy, thereby improving survival outcomes in orthotopic GSC-derived and patient-derived GBM xenograft models. Conclusions: This study underscores the pivotal role of MYC-mediated PHGDH activation in driving GSC malignant progression and radioresistance in GBM. Targeting PHGDH presents a promising approach to enhance radiotherapy efficacy in GBM patients.